University of Saint Louis Tuguegarao
Basic Education School
Names:
1. Keynes Joven C. Grande
2. Sergei Frahnielzo M. Rodriquez
3. Kyla Fhaye Fronda Martinez
4. Rihanna Riezza C. Ramos
5. Rose Angel L. Rementilla
Research Title
Experimental Investigations into the
Insecticidal, Fungicidal, and
Bactericidal Properties of Pyrolysis
Bio-oil from Tobacco Leaves Using
a Fluidized Bed Pilot Plant
Senior High School Unit
Practical Research 1
Literature Review (Abstract) Type of Research (explanation) Research Gap
Applied
Tobacco bio-oil, gases, and char Applied
were produced through pyrolysis of
tobacco leaves using a fluidized
bed pilot plant under varying
temperature (350, 400, 450, 500,
550, and 600 °C) and residence
time (5, 10, and 17 s) conditions.
The optimized condition for the
production of bio-oil was found to
be at 500 °C at a vapor residence
time of 5 s, giving a bio-oil yield of
43.4%. The Colorado Potato Beetle
(CPB) Leptinotarsa
decemlineata L. (Coleoptera:
Chrysomelidae), a destructive pest
toward potato crops, and three
microorganisms (Streptomyces
scabies, Clavibacter
michiganensis, and Pythium
ultimum), all problematic in
Canadian agriculture, were strongly
affected by tobacco bio-oil
generated at all pyrolysis
temperatures. Nicotine-free

In vitro antibacterial effect of
tobacco leaf exudates against two
bacterial plant pathogens
fractions of the tobacco bio-oil were
prepared through liquid−liquid
extraction, and high mortality rates
for the CPB and inhibited growth
for the microorganisms were still
observed. A potential pesticide from
tobacco bio-oil adds value to the
biomass as well as the pyrolysis
process.
Natural products are an alternative Applied
to control microorganisms that
cause diseases in crops. This work
aimed to evaluate different solvents
for obtaining crude extracts from
tobacco leaf exudates and to
determine in vitro effect of these
extracts against two
phytopathogenic
bacteria: Xanthomonas campestris
(Xc) and Pectobacterium
carotovorum (Pc). Crude extracts
from ten tobacco lines using
solvents with polarities between 3.1
and 6.2 (dichloromethane, n-
butanol, ethyl acetate, methanol
and ethanol 90%) were obtained.
Ethanol 90% was selected as the
best solvent for obtaining extracts
from tobacco leaf exudates and as
a substitute of dichloromethane
due to the best yield. The chemical
composition diversity of the
ethanolic extracts was revealed by
thin-layer chromatography. The
antibacterial activity was evaluated
by agar disk diffusion method
recording the inhibition zones.
Growth inhibition was observed for
all extracts against Xc, and the
better activity corresponded to the
lines Nic 1061"TI 1738" and Nic
1016 "Incekara" until a minimal

Tobacco Extract-Based
Biopesticide from Cigarette Butt
Waste for Corn plants (Zea mays
L.)
amount of 5 μg/ disc, with higher
yield in case of the line Nic1061 .
Only the extract of the line Nic
1015 was able to inhibit the growth
of Pc until a minimal inhibitory
concentration of 5 μg/disc. These
results suggest a potential use of
crude extracts from lines Nic 1061
and Nic 1015 "TI 1341" as an
effective agent for the crop
protection
against Xc and Pc respectively.
Cigarette butt waste can be used Applied
for biopesticides for
environmentally friendly agricultural
activities.
Biopesticides cigarette can be
produced by conducting a
maceration extraction test using
polar, non-polar, and
semi-polar compounds and
conducting a Mass Spectrometer
Chromatography (GC-MS) test.
This study aims to
determine the effectiveness of bio-
pesticide based on tobacco extract
from cigarette butt waste for maize
(Zea
mays L.)". The highest yield value
was the ethanol sample of
1.208%, compared to the N-
hexane sample of
0.25% and the DCM sample of
0.6%. For the comparison of
solvents used were 500 ml of
ethanol, 250 ml of N-
hexane, and 250 ml for DCM as
well. After conducting the GC-MS
test, the compounds obtained are
alkaloid
compounds, phenolic compounds,
sesquiterpenoid compounds, and

The Effectiveness of Tobacco Oil
Fungicidal Preparation Towards
Powdery Mildew of Wheat
(Blumeria graminis f. sp. tritici)
Extract of Nicotiana tabacum as a
potential control agent
of Grapholita molesta (Lepidoptera:
Tortricidae)
terpenoid compounds, each of
which has
benefits as a pest control or
deterrent to plants. For the
effectiveness test results on Corn
(Zea mays L.), there
was no intensity of pest attack on
the ethanol sample, N-hexane
sample, and DCM sample. In
contrast, there
was a pest attack intensity of 0.3%
for the control sample. Therefore,
the Biopesticides studied are
similar to
synthetic pesticides because they
can eradicate pests and make
growth effective faste
The pesticide formulation prepared Applied
as "ready to be used plant
protection product" on the basis of
tobacco oil was created and tested
in two years field trials against
powdery mildew of wheat
(Blumeria graminis f. sp. tritici). The
results showed strong fungicidal
effect without any phytotoxic
activities (chemotherapeutic
coefficient = 10) onto treated
plants. This indicates the future
development of this pesticide
formulation as effective, natural
friendly and cheap product for plant
protection, which can be used both
in commercial and organic
agriculture.
Oriental fruit moth, Grapholita Applied
molesta (Busck) (Lepidoptera:
Tortricidae), is an important pest of
stone and pome fruits. Growers
usually depend on chemical
insecticides to control this pest, but
demand for more environmentally-
friendly means of controlling pests
is increasing. At least 91 plant
extracts have been reported to be
effective against other
lepidopterans, but their acute
toxicity against G. molesta has
rarely been studied. Among these
91 materials, we assessed the
residual toxicity of 32 extracts
against first instar larvae (< 5 h old)
of G. molesta in the
laboratory. Nicotiana tabacum L.,
used at the concentration of 2
mg/ml, showed the highest
corrected mortality (92.0%) with a
lethal time (LT50) value of 12.9 h.
The extract was followed in its
efficacy by Allium sativum L.
(88.0%), Zanthoxylum
piperitum (L.) De Candolle (70.0%),
and Sapindus mukorossi Gaertner
(65.0%), when mortality was
assessed at 20 h after exposure.
Against adult fruit moths (< 5 d
old), N. tabacum also showed the
highest corrected mortality among
tested extracts, being 85 and 100%
in adult females and males,
respectively, at 168 h after
exposure. However, there was no
synergistic effect of the combined
application of any of the top four
extracts in either laboratory or
greenhouse assays. Oviposition
by G. molesta on peach twigs was
reduced 85–90%
when N. tabacum was applied at 4
ml/ twig compared to control
(methanol), demonstrating
that N. tabacum may have potential
for use as a botanical insecticide

Antimicrobial Activities of Extracts
of Tobacco Leaf (Nicotiana
tabacum) and Its Grounded Snuff
(Utaba) on Candida albicans and
Streptococcus pyogenes
against G. molesta.
This study was designed to Applied
investigate the antimicrobial activity
of leaf and ground snuff extracts of
Tobacco (Nicotiana tabacum)
against Candida albicans and
Streptococcus pyogenes using
methanol and water as extracting
solvents. The study employed the
agar diffusion and tube dilution
assays. Methanol extracts of
tobacco leaf produced zones of
inhibition of 13.0 mm against
Streptococcus and 9.5 mm against
Candida, whereas the water
extracts produced inhibition zones
of 10.0 mm for Streptococcus and
no inhibitory activity on Candida. A
minimum inhibitory concentration of
25 mg/ml was recorded by the
methanol extracts of tobacco
leaves against Candida and 100
mg/ml MIC against Streptococcus.
The methanolic leaf extracts had
both bactericidal and fungicidal
effect on both Streptococcus and
Candida at a concentration of 200
mg/ml. The zones of inhibition
obtained from methanolic extracts
of grounded snuff against
Streptococcus was 10.5 mm and
15.0 mm against Candida whereas
the water extracts produced
inhibition zones of 7.5 mm for
Streptococcus and 11.0 mm
against Candida. A minimum
inhibitory concentration of 100
mg/ml was recorded by both
methanolic and water snuff extracts
against Streptococcus. While the
MIC obtained from the methanolic
extract of ground snuff against

Chemical Constituents
from Nicotiana tabacum L. and
Their Antifungal Activity
Candida was 50 mg/ml. The water
extracts of ground snuff showed no
bactericidal or fungicidal activity.
Whereas 200 mg/ml of the
methanolic extract of ground snuff
was microbiocidal against
Streptococcus and Candida. In
summary, the study showed that
Grounded snuff is more of an
antifungal agent than antibacterial
while tobacco leaves have great
antibacterial potential. This may
justify the use of tobacco leaves
and its ground snuff in the
treatment of oral thrush caused by
Candida albicans and strep throat
caused by Streptococcus
pyogenes.
A phytochemical investigation of an Applied
aqueous ethyl alcohol (EtOH)
extract of the flowers of Nicotiana
tabacum L. led to the identification
of one new eudesmane-type
sesquiterpenoid (1) and 14 known
compounds, including 7 flavones
(2-8), 6 phenolic acids (9-14), and
1 coumarin (15). The chemical
structure of 1 was established on
the basis of high-resolution
electrospray ionization (HRESIMS)
and 1D and 2D NMR analyses,
while the other structures were
elucidated by 1D NMR and LC–MS
data. The antiphytopathogenic
activities of all the isolated
compounds were tested to
determine their antifungal activity
against Valsa
mali var. mali, Alternaria porri,
and Botrytis cinerea at a
concentration of 10 µg/mL.
Compound 4 exhibited more potent

Computational and experimental
investigation of antibacterial and
antifungal properties of Nicotiana
tabacum extracts
antifungal effects against Valsa
mali var. mali with a % growth
inhibition of 47.9 ± 1.2%, while the
others only showed insignificant
activities.
The identification of novel anti- Applied
infective agents of synthetic and
natural origin represents one of the
main aims of contemporary drug
discovery. In the current work, four
different varieties of Nicotiana
tabacum, namely, K399, SPG28,
Swat No. 1, and Swat No. 2, were
studied to assess the antibacterial
and antifungal properties of their
extracts. The extracts contain
anthraquinones, alkaloids,
saponins, terpenoids, tannins,
resins, steroids, proteins, and
carbohydrates, and the
antibacterial and antifungal
activities were evaluated toward
four bacterial and four fungal
strains. N. tabacum K399 showed
the highest zone of inhibition
against E. coli. Similarly, K399
showed the highest antifungal
potential, as the highest zone of
inhibition for the set was detected
against C. albicans. Then, the
underlying molecular mechanism
was further investigated, and the
extracts were tested for their
inhibitory potential against urease,
an enzyme which is conserved in
bacteria and fungi. Additionally,
computational tools were enrolled
to assess the role of rutin and
chlorogenic acid, which are among
the main constituents of N.
tabacum leaves, in interacting with
urease through molecular docking.

Antifungal properties of leaf extract
of neem and tobacco on three
fungal pathogens of tomato
(Lycopersicon Esculentum Mill)
Combined together, the
computational and experimental
results support the antibacterial
and antifungal potential of N.
tabacum extracts, particularly, that
obtained from K399 variety.
Laboratory studies were carried out Applied
to isolate, identify and control the
fungi associated with fruit rot of
tomato (Lycopersicon esculentum).
Aspergillus viridae, Penicillium
digitatum and Rhizopus sp were
isolated from samples collected
from Lokoja, Kabba, Okene,
Anyigba and Ankpa, all in Kogi
State, Nigeria. Plant extracts
employed to control the vegetative
mycelial growth were from neem
(Azadirachta indica) and tobacco
(Nicotiana tabacum) at 20%, 30%,
40%, 50% and 60%
concentrations. In vitro application
of extracts for the control showed
that tobacco (Nicotiana tabacum)
had fungitoxic effect that controlled
the mycelia growth; there was
complete inhibition (0.00 + 0.00) at
60% on Aspergillus viridae and
Penicillium digitatum. Tobacco
extract on Rhizopus showed
progressive retardations on the
vegetative growth. Vegetative
growth of the fungi on neem extract
decreased with increase in
concentrations; with its effect
(extract) being felt more on
Penicillium digitatum at 60%
concentration (0.41 + 0.55)
when compare with the control.
The inhibitory action of the two
extracts on mycelial growth
increased with increase in

Bioenergy II: Characterization of the
Pesticide Properties of Tobacco Bio-Oil
concentrations; giving toxicity
profile of 60% > 50% in that order,
and were significant at 0.05%.
Bio-oil from tobacco plants is known
to contain the highly toxic chemical
nicotine (Scott D.S., 1997) (oral LD50
of 50 mg/kg for rats). In addition to its
use in pharmaceuticals, nicotine is
permitted as a pesticide in Canada,
although its use as a pesticide is
limited because of its extreme toxicity
to and dermal absorption for
mammals. Thus, it was expected that
tobacco bio-oil would have pesticide
properties due to the presence of
nicotine. It was found in our research
group, however, that nicotine-free
fractions of tobacco bio-oil were lethal
to the Colorado Potato Beetle (CPB,
Leptinotarsa decemlineata), as well as
successful at inhibiting the growth of
two bacteria (Clavibacter
michiganensis subsp. Booker et al.:
Characterization of the Pesticide
Properties of Tobacco Bio-Oil 1
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and Streptomyces scabies) and a
fungus (Pythium ultimum) (Bedmutha,
2008, Booker, et al., 2008). The activity
of the tobacco bio-oil towards the
microorganisms will be discussed in
this paper. S. scabies disfigures potato
crops making them unmarketable, C.
michiganensis seriously disfigures or
kills tomato plants, and P. ultimum
causes seedling damping off in a
variety of vegetable plants. Tobacco
bio-oil’s pesticide properties are an
exciting and potentially economically
valuable finding. Although the broad,
general composition of bio-oil is
known (mixture of alcohols, phenols,
lignin, etc), no report, to our
knowledge, has clearly identified
which of these chemicals are
responsible for the pesticide
characteristics in biooil. Phenolic
compounds have been assumed
responsible for the observed
antifungal activity in wood bio-oil, but
this correlation has not been
experimentally supported. While some
of the identified chemicals in bio-oil
are known to be active towards pests
(ie. o-cresol, m-cresol, and p-cresol) (EI
DuPont de Numours & Co., 1983), the
concentrations at which they are
found in bio-oil may not be sufficient
to account for the observed activity.